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MGMT promoter methylation in gliomas-assessment by pyrosequencing and quantitative methylation-specific PCR

BACKGROUND: Methylation of the O(6)-methylguanine-DNA methyltransferase (MGMT) gene promoter is a favorable prognostic factor in glioblastoma patients. However, reported methylation frequencies vary significantly partly due to lack of consensus in the choice of analytical method. METHOD: We examined...

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Autores principales: Håvik, Annette Bentsen, Brandal, Petter, Honne, Hilde, Dahlback, Hanne-Sofie Spenning, Scheie, David, Hektoen, Merete, Meling, Torstein Ragnar, Helseth, Eirik, Heim, Sverre, Lothe, Ragnhild A, Lind, Guro Elisabeth
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2012
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3311573/
https://www.ncbi.nlm.nih.gov/pubmed/22390413
http://dx.doi.org/10.1186/1479-5876-10-36
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author Håvik, Annette Bentsen
Brandal, Petter
Honne, Hilde
Dahlback, Hanne-Sofie Spenning
Scheie, David
Hektoen, Merete
Meling, Torstein Ragnar
Helseth, Eirik
Heim, Sverre
Lothe, Ragnhild A
Lind, Guro Elisabeth
author_facet Håvik, Annette Bentsen
Brandal, Petter
Honne, Hilde
Dahlback, Hanne-Sofie Spenning
Scheie, David
Hektoen, Merete
Meling, Torstein Ragnar
Helseth, Eirik
Heim, Sverre
Lothe, Ragnhild A
Lind, Guro Elisabeth
author_sort Håvik, Annette Bentsen
collection PubMed
description BACKGROUND: Methylation of the O(6)-methylguanine-DNA methyltransferase (MGMT) gene promoter is a favorable prognostic factor in glioblastoma patients. However, reported methylation frequencies vary significantly partly due to lack of consensus in the choice of analytical method. METHOD: We examined 35 low- and 99 high-grade gliomas using quantitative methylation specific PCR (qMSP) and pyrosequencing. Gene expression level of MGMT was analyzed by RT-PCR. RESULTS: When examined by qMSP, 26% of low-grade and 37% of high-grade gliomas were found to be methylated, whereas 97% of low-grade and 55% of high-grade gliomas were found methylated by pyrosequencing. The average MGMT gene expression level was significantly lower in the group of patients with a methylated promoter independent of method used for methylation detection. Primary glioblastoma patients with a methylated MGMT promoter (as evaluated by both methylation detection methods) had approximately 5 months longer median survival compared to patients with an unmethylated promoter (log-rank test; pyrosequencing P = .02, qMSP P = .06). One third of the analyzed samples had conflicting methylation results when comparing the data from the qMSP and pyrosequencing. The overall survival analysis shows that these patients have an intermediate prognosis between the groups with concordant MGMT promoter methylation results when comparing the two methods. CONCLUSION: In our opinion, MGMT promoter methylation analysis gives sufficient prognostic information to merit its inclusion in the standard management of patients with high-grade gliomas, and in this study pyrosequencing came across as the better analytical method.
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spelling pubmed-33115732012-03-24 MGMT promoter methylation in gliomas-assessment by pyrosequencing and quantitative methylation-specific PCR Håvik, Annette Bentsen Brandal, Petter Honne, Hilde Dahlback, Hanne-Sofie Spenning Scheie, David Hektoen, Merete Meling, Torstein Ragnar Helseth, Eirik Heim, Sverre Lothe, Ragnhild A Lind, Guro Elisabeth J Transl Med Research BACKGROUND: Methylation of the O(6)-methylguanine-DNA methyltransferase (MGMT) gene promoter is a favorable prognostic factor in glioblastoma patients. However, reported methylation frequencies vary significantly partly due to lack of consensus in the choice of analytical method. METHOD: We examined 35 low- and 99 high-grade gliomas using quantitative methylation specific PCR (qMSP) and pyrosequencing. Gene expression level of MGMT was analyzed by RT-PCR. RESULTS: When examined by qMSP, 26% of low-grade and 37% of high-grade gliomas were found to be methylated, whereas 97% of low-grade and 55% of high-grade gliomas were found methylated by pyrosequencing. The average MGMT gene expression level was significantly lower in the group of patients with a methylated promoter independent of method used for methylation detection. Primary glioblastoma patients with a methylated MGMT promoter (as evaluated by both methylation detection methods) had approximately 5 months longer median survival compared to patients with an unmethylated promoter (log-rank test; pyrosequencing P = .02, qMSP P = .06). One third of the analyzed samples had conflicting methylation results when comparing the data from the qMSP and pyrosequencing. The overall survival analysis shows that these patients have an intermediate prognosis between the groups with concordant MGMT promoter methylation results when comparing the two methods. CONCLUSION: In our opinion, MGMT promoter methylation analysis gives sufficient prognostic information to merit its inclusion in the standard management of patients with high-grade gliomas, and in this study pyrosequencing came across as the better analytical method. BioMed Central 2012-03-06 /pmc/articles/PMC3311573/ /pubmed/22390413 http://dx.doi.org/10.1186/1479-5876-10-36 Text en Copyright ©2012 Håvik et al; licensee BioMed Central Ltd. http://creativecommons.org/licenses/by/2.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research
Håvik, Annette Bentsen
Brandal, Petter
Honne, Hilde
Dahlback, Hanne-Sofie Spenning
Scheie, David
Hektoen, Merete
Meling, Torstein Ragnar
Helseth, Eirik
Heim, Sverre
Lothe, Ragnhild A
Lind, Guro Elisabeth
MGMT promoter methylation in gliomas-assessment by pyrosequencing and quantitative methylation-specific PCR
title MGMT promoter methylation in gliomas-assessment by pyrosequencing and quantitative methylation-specific PCR
title_full MGMT promoter methylation in gliomas-assessment by pyrosequencing and quantitative methylation-specific PCR
title_fullStr MGMT promoter methylation in gliomas-assessment by pyrosequencing and quantitative methylation-specific PCR
title_full_unstemmed MGMT promoter methylation in gliomas-assessment by pyrosequencing and quantitative methylation-specific PCR
title_short MGMT promoter methylation in gliomas-assessment by pyrosequencing and quantitative methylation-specific PCR
title_sort mgmt promoter methylation in gliomas-assessment by pyrosequencing and quantitative methylation-specific pcr
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3311573/
https://www.ncbi.nlm.nih.gov/pubmed/22390413
http://dx.doi.org/10.1186/1479-5876-10-36
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